Diazacycloalkanedione derivatives

ABSTRACT

Compounds of formula I  
                 
 
     wherein  
     R is carboxy, esterified carboxy or amidated carboxy;  
     R 1  and R 3  are independently lower alkyl, (hydroxy, lower alkoxy, amino, acylamino, mono- or di-lower alkylamino or mercapto)-lower alkyl, lower alkyl-(thio, sulfinyl or sulfonyl)-lower alkyl, cycloalkyl, aryl, biaryl, (cycloalkyl, aryl or biaryl)-lower alkyl, or (carboxy, esterified carboxy or amidated carboxy)-lower alkyl;  
     R 2  is hydrogen, lower alkyl, cycloalkyl, aryl, biaryl, arylaminocarbonyl, or aryl-(oxy, thio or amino);  
     n is 1 or 2;  
     Y is lower-alkylene or lower alkenylene;  
     and pharmaceutically acceptable salts thereof; which are useful as LFA-1 antagonists.

CROSS-REFERENCE TO RELATED APPLICATIONS

[0001] This application claims the benefit of provisional ApplicationNo. 60/237,231 filed Oct. 2, 2000, which is incorporated herein byreference.

BACKGROUND OF THE INVENTION

[0002] Lymphocyte function associated antigen 1 (LFA-1) mediateslymphocyte adhesion and plays a key role in inflammation and the immuneresponse.

[0003] LFA-1 binds to intracellular adhesion molecules, e.g., ICAM-1,ICAM-2 or ICAM-3. The LFA-1/ICAM-1, ICAM-2 or ICAM-3 mediatedinteractions have been implicated in various disorders includingtransplant rejection, chronic inflammation, psoriasis,eczema/dermatitis, asthma and arthritis. LFA-1 antagonists are thususeful for the treatment and/or prevention of disorders which areresponsive to LFA-1 inhibition, e.g., those cited herein, such asrheumatoid arthritis.

SUMMARY OF THE INVENTION

[0004] The present invention relates to diazacycloalkanedionederivatives, e.g., diazepanedione (hexahydrodiazepinedione) anddiketopiperazine derivatives of formula I described herein which areparticularly useful as LFA-1 antagonists, pharmaceutical compositionsthereof, their methods of preparation and methods of treating conditionsin mammals which are responsive to LFA-1 antagonism using said compoundsor pharmaceutical compositions thereof.

DETAILED DESCRIPTION OF THE INVENTION

[0005] The invention relates particularly to compounds of formula I

[0006] wherein

[0007] R is carboxy, esterified carboxy or amidated carboxy;

[0008] R₁ and R₃ are independently lower alkyl, (hydroxy, lower alkoxy,amino, acylamino, mono- or di-lower alkylamino or mercapto)-lower alkyl,lower alkyl-(thio, sulfinyl or sulfonyl)-lower alkyl, cycloalkyl, aryl,biaryl, (cycloalkyl, aryl or biaryl)-lower alkyl, or (carboxy,esterified carboxy or amidated carboxy)-lower alkyl;

[0009] R₂ is hydrogen, lower alkyl, cycloalkyl, aryl, biaryl,arylaminocarbonyl, or aryl-(oxy, thio or amino); n is 1 or 2;

[0010] Y is lower-alkylene or lower alkenylene;

[0011] and pharmaceutically acceptable salts thereof.

[0012] A particular embodiment of the invention relates to the compoundsof formula I wherein R is carboxy derivatized in form of apharmaceutically acceptable amide; R₁ is aryl-lower alkyl; R₂ is aryl;R₃ is lower alkyl, aryl, or cycloalkyl-lower alkyl; and Y isC₁-C₄-alkylene or C₂₋₄-alkenylene; and pharmaceutically acceptable saltsthereof.

[0013] A more particular embodiment of the invention relates to thecompounds of formula II

[0014] wherein R₁′ is bicyclic aryl-lower alkyl; R₂′ is bicyclic aryl;R₃′ is monocyclic aryl or lower alkyl; R₄ is hydrogen or lower alkyl; nis 1 or 2; Y is C₁₋₄-alkylene; and pharmaceutically acceptable saltsthereof.

[0015] Particular embodiments of the invention relate to compounds offormula I and II wherein n is 1 and wherein n is 2.

[0016] Further preferred are said compound of formula II wherein R₁′ isnaphthyl-lower alkyl or quinolinyl-lower alkyl; R₂′ is quinolinyl; R₃′is isobutyl; Y is methylene, and n is 2; and pharmaceutically acceptablesalts thereof.

[0017] Compounds of the invention may possess one or more asymmetriccenters and can exist as diastereomers, racemates and the enantiomersthereof, all of which are within the purview of the invention.

[0018] Preferred are the diastereomers of the formula Ia or IIa

[0019] wherein n, Y, R, R₁, R₂ and R₃ in formula Ia, and n, Y, R₁′, R₂′,R₃′ and R₄ in formula IIa have meaning as defined above.

[0020] Unless otherwise indicated, the general definitions used hereinhave the following meaning within the scope of the present invention.

[0021] Aryl represents carbocyclic or heterocyclic aryl, eithermonocyclic or bicyclic.

[0022] Monocyclic carbocyclic aryl represents optionally substitutedphenyl, being preferably phenyl or phenyl substituted by one to threesubstituents, such being advantageously lower alkyl, hydroxy, loweralkoxy, acyloxy, halogen, cyano, trifluoromethyl, carbocyclic aryloxy orcarbocyclic aryl-lower alkoxy; or phenyl substituted on adjacent carbonatoms by lower alkylene or by lower alkylene interrupted by O, or S orby N optionally substituted by lower alkyl.

[0023] Bicyclic carbocyclic aryl represents 1- or 2-naphthyl or 1- or2-naphthyl substituted by, e.g., lower alkyl, lower alkoxy or halogen,advantageously 2-naphthyl. Naphthyl may also be named naphthalenyl.

[0024] Monocyclic heterocyclic aryl represents, e.g., optionallysubstituted thiazolyl, thienyl, furanyl or pyridyl.

[0025] Optionally substituted furanyl represents 2- or 3-furanylpreferably substituted by lower alkyl.

[0026] Optionally substituted pyridyl represents 2-, 3- or 4-pyridyl or2-, 3- or 4-pyridyl preferably substituted by lower alkyl, halogen orcyano.

[0027] Optionally substituted thienyl represents 2- or 3-thienyl or 2-or 3-thienyl preferably substituted by lower alkyl.

[0028] Optionally substituted thiazolyl represents, e.g., 2- or4-thiazolyl, or 2- or 4-thiazolyl preferably substituted by lower alkyl.

[0029] Bicyclic heterocyclic aryl represents, e.g., quinolinyl,isoquinolinyl, indolyl or benzothiazolyl optionally substituted byhydroxy, lower alkyl, lower alkoxy or halogen. Indolyl may also bear asubstituent attached to the ring nitrogen, e.g., lower alkyl oraryl-lower alkyl.

[0030] Aryl-lower alkyl is advantageously arylmethyl optionallysubstituted on phenyl by one or two of lower alkyl, lower alkoxy,hydroxy, lower alkanoyloxy, halogen, cyano or trifluoromethyl.

[0031] Biaryl represents phenyl substituted by carbocyclic aryl orheterocyclic aryl as defined herein, ortho, meta or para to the point ofattachment of the phenyl ring, advantageously para, such as biphenyl,particularly 4-biphenyl, or pyridylphenyl, particularly 4-pyridylphenyl.

[0032] The term “lower” referred to herein in connection with organicradicals or compounds respectively defines such with up to and including7, preferably up and including 4 and advantageously one or two carbonatoms. Such may be straight chain or branched.

[0033] A lower alkyl group preferably contains 1-4 carbon atoms andrepresents for example methyl, ethyl, propyl or butyl.

[0034] A lower alkylene group preferably contains 1-4 carbon atoms, andrepresents, for example, methylene, ethylene, propylene and the like.

[0035] A lower alkenylene group preferably contains 2-4 carbon atoms,and represents, for example, propenylene.

[0036] Cycloalkyl represents preferably cyclopentyl, cyclohexyl orcycloheptyl.

[0037] A lower alkoxy group preferably contains 1-4 carbon atoms andrepresents for example, methoxy, propoxy, isoproproxy or advantageouslyethoxy.

[0038] Halogen (halo) preferably represents fluoro or chloro, but mayalso be bromo or iodo.

[0039] Acyl is derived from a carboxylic acid or carbonic acid andrepresents preferably optionally substituted lower alkanoyl, aroyl,lower alkoxycarbonyl or aryl-lower alkoxycarbonyl, advantageously aroyl.

[0040] Lower alkanoyl is preferably acetyl, propionyl, butyryl, orpivaloyl.

[0041] Optionally substituted lower alkanoyl for example representslower alkanoyl or lower alkanoyl substituted, e.g., by loweralkoxycarbonyl, lower alkanoyloxy, lower alkanoylthio, lower alkoxy, orby lower alkylthio.

[0042] Aroyl is preferably monocyclic carbocyclic or monocyclicheterocyclic aroyl.

[0043] Monocyclic carbocyclic aroyl is preferably benzoyl or benzoylsubstituted by lower alkyl, lower alkoxy, halogen or trifluoromethyl.

[0044] Monocyclic heterocyclic aroyl is preferably pyridylcarbonyl orthienylcarbonyl.

[0045] Acyloxy is preferably optionally substituted lower alkanoyloxy,lower alkoxycarbonyloxy, monocyclic carbocyclic aroyloxy or monocyclicheterocyclic aroyloxy.

[0046] Esterified carboxy represents carboxy derivatized in form of apharmaceutically acceptable ester, preferably lower alkyl esters,cycloalkyl esters, lower alkenyl esters, benzyl esters, mono ordisubstituted lower alkyl esters, e.g., the ω-(amino, mono- or di-loweralkylamino, carboxy, lower alkoxycarbonyl)-lower alkyl esters, theα-(lower alkanoyloxy, lower alkoxycarbonyl or di-loweralkylaminocarbonyl)-lower alkyl esters, such as the pivaloyloxy-methylester, and the like conventionally used in the art.

[0047] Amidated carboxy represents carboxy derivatized in form of apharmaceutically acceptable amide, e.g., primary, secondary and tertiaryamides, e.g., the unsubstituted the N-mono-lower alkyl or N,N-di-loweralkylamides, or the amides of cyclic amines, e.g., of piperidine,pyrrolidine, morpholine or optionally substituted piperazine.

[0048] Pharmaceutically acceptable salts of acids of the invention aresalts derived from pharmaceutically acceptable bases, e.g., alkali metalsalts (e.g., sodium, potassium salts), alkaline earth metal salts (e.g.,magnesium, calcium salts), amine salts (e.g., ethanolamine,diethanolamine, lysine and tromethamine salts) and the likeconventionally used in the art.

[0049] Pharmaceutically acceptable salts of basic compounds of theinvention are acid addition salts, e.g., of mineral acids, organiccarboxylic acids, and organic sulfonic acids, e.g., hydrochloric acid,methanesulfonic acid, and the like.

[0050] The compounds of the invention of formula I can be prepared

[0051] (a) by cyclizing a compound of the formula III or a reactivefunctional derivative thereof

[0052] wherein R, R₁-R₃, Y and n have meaning as defined above; or

[0053] (b) by cyclizing a compound of the formula IV or a reactivefunctional derivative thereof

[0054] wherein R, R₁-R₃, Y and n have meaning as defined above.

[0055] Abbreviations used throughout the application are those commonlyused in the art, e.g.,

[0056] DCCI=Dicyclohexylcarbodiimide

[0057] DIPCI=Diisopropylcarbodiimide

[0058] HOBT=1-Hydroxybenzotriazole

[0059] NMM=N-methylmorpholine

[0060] DIEA=Diisopropylethylamine

[0061] HATU=O-(7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluroniumhexafluorophosphate

[0062] r.t.=room temperature

[0063] TEA=Triethylamine

[0064] TFA=trifluoroacetic acid

[0065] A reactive functional derivative of a compound of formula III orIV includes, e.g., a halide, ester or anhydride of the respectivecarboxylic acid.

[0066] The cyclization of an intermediate of the formula III or IV maybe carried out according to methodology known in the art for formationof a lactam, e.g., using a condensing agent, such as HATU in thepresence of diisopropylethylamine in a polar solvent, such asdimethylformamide.

[0067] Illustrative of the invention, the preparation of compounds offormula II wherein n is 1 can be carried out as illustrated below and inthe examples herein.

[0068] Further illustrative of the invention, the preparation ofcompounds of formula II wherein n is 2 can be carried out as illustratedbelow and in the examples herein.

[0069] The preparation of corresponding compounds of compounds offormula IIa can be carried out according to the above synthetic schemesusing starting materials with appropriate configurations at theasymmetric centers bearing the R₁′ and R₃′ substituents, e.g., asillustrated in the examples.

[0070] The compounds of the invention, in free form or inpharmaceutically acceptable salt form exhibit valuable pharmacologicalproperties, e.g., as LFA-1 antagonists inhibiting LFA-1/ICAM-1, ICAM-2or ICAM-3 interactions or inhibiting inflammation, e.g., as determinedin in vitro and in vivo tests and are therefore indicated for therapy inthe treatment of disorders responsive to LFA-1 inhibition.

[0071] The pharmacological properties can be demonstrated as follows:

[0072] A. In Vitro: Cell Free Assay

[0073] The assay measures the binding of soluble human ICAM-1 toimmobilized human LFA-1. LFA-1 is purified from JY cells, a humanlymphoblastoid B cell-line, by immunoaffinity chromatography asdescribed by Dustin et al. (J. Immunol. 148, 2654-2663, 1992). ICAM-1mouse Cκ fusion protein (ICAM-1) is produced using the baculovirussystem as described by Weitz-Schmidt et al. (Anal. Biochem.238,184-190,1996).

[0074] Purified LFA-1 is diluted 1:20 in phosphate buffered saline (PBS)containing 2 mM MgCl₂, pH 7.4 and coated onto microtitre plates (Nunc)at 37° C. for 3 h. Plates are blocked with 1% heat-treated BSA in PBSfor 2 hours at 37° C. followed by a washing step using PBS, 2 mM MgCl₂,1% fetal calf serum, pH 7.4 (assay buffer). Compounds dissolved at 10 mMin DMSO are diluted in assay buffer and added to the plates.Biotinylated recombinant ICAM-1 in assay buffer (6 μg/ml) is added andallowed to bind at 37° C. for one hour. After incubation, wells arewashed with assay buffer. Streptavidin-peroxidase diluted 1:5000 inassay buffer is added and incubated for 45 min at 37° C. Plates are thenwashed with assay buffer and 2,2′-azino-bis(3-ethylbenzothiazoline-6sulfonic acid) diammonium salt substrate solution is added to each well.The reaction is stopped after 20 min and bound ICAM-1 is determined bymeasuring the optical density at 405 nm in a microplate reader.

[0075] In this assay, compounds of the invention inhibit adhesion ofLFA-1 to ICAM-1. Compounds of Examples 1 and 4 have an IC₅₀ of about 30nM and 23 nM, respectively, in this assay.

[0076] B. In Vivo Assays

[0077] i) Murine Thioglycollate Induced Peritonitis

[0078] Thioglycollate is injected i.p. to mice and immediatelythereafter the compound to be tested is given s.c. or p.o. The mice aresacrificed after 4 hours, the peritoneal cavity lavaged and total numberof neutrophils in the lavage fluid is determined.

[0079] In this assay, the compounds of the invention inhibitthioglycollate induced neutrophil migration when administered s.c. orp.o. at a dose of about 0.01-100 mg/kg, either at the time of thethioglycollate injection or 3 hours before.

[0080] ii) Allergic Contact Dermatitis (ACD)

[0081] Groups of 8 female NMRI mice are sensitized on the shaved abdomenwith 50 μl of oxazolone (Sigma, 2% in acetone) and challenged with 10 μlof 0.2 or 2.0% oxazolone on the inner surface of the right ear 7 dayslater. The low concentration of oxazolone for induction of theelicitation phase is used for testing compounds on systemic activitywhereas the high concentration is applied for systemic testing. Theunchallenged left ears serve as normal controls and dermatitis isevaluated from the individual differences in pinnal weight, which istaken as a measure of increase in inflammatory swelling 24 h after thechallenge. Dermatitis is evaluated in test groups and for comparison incontrol groups. The test groups are treated with the test compoundseither orally (twice, 2 h and immediately before challenge),subcutaneously (immediately before challenge) or topically (30 min afterchallenge at the site of elicitation of the ACD); the controls aretreated similarly with the vehicles alone. For oral and subcutaneousadministration the compounds are administered in an oil in wateremulsion, for topical administration the compounds are administered in amixture of ethanol, acetone and dimethylacetamide. The data of the test-and the vehicle-treated control groups are statistically analysed byANOVA followed by Dunnet T-test (normal distribution or data) or by Hand U-test, respectively. When administered p.o. at a dose of from about0.1 to 20 mg/kg, compounds of the invention inhibit the elicitationphase of allergic contact dermatitis.

[0082] Antiinflammatory, antiarthritic and immunosuppressant activitycan be determined according to tests well known in the art, e.g., in thecarrageenan test, the antigen-induced arthritis test and experimentalallergic encephalitis test.

[0083] iii) Rat Carrageenan Edema Model.

[0084] Male OFA rats are treated orally with the compound to be testedor vehicle (saline). One to three hours later (0 hours) the rats receivea 100 μl intra-plantar injection of a 1% w/v carrageenan solution in0.9% saline in the hind paw and the diameter of the paw is measured bymeans of a micro-calliper. The paw diameter measurements are repeated attimes +3 and +5 hours after injection of the carrageenan. Percentageinhibition of paw swelling at 3 and 5 hours are calculated by referenceto vehicle treated animals (0% inhibition).

[0085] Illustrative of the invention, the compound of example 1 inhibitspaw swelling in the rat carrageenan edema model at a dose range of about0.3 to 30 mg/kg p.o.

[0086] iv) Mouse Antigen-induced Arthritis Model.

[0087] The test for rheumatoid arthritis is carried out by amodification of that described by Van de Loo et al., Arthritis Rheum.1995; 35:164-172, e.g., as follows:

[0088] Female OFA-1 mice are sensitised intradermally on the back at twosites to methylated bovine serum albumin (mBSA) homogenised 1:1 withcomplete Freund's adjuvant on days −21 and −14 (0.1 ml containing 1mg/ml mBSA). On day 0, the right knee receives 10 μl of 10 mg/ml mBSA in5% glucose solution (antigen injected knee), while the left kneereceives 10 μl of 5% glucose solution alone (vehicle injected knee). Thediameters of the left and right knees are then measured using calipersimmediately after the intra-articular injections and again on days 2, 4,7, 9, 11 and 14. Compounds to be tested are administered once or twicedaily by oral gavage; vehicle control (saline) is administered at 5ml/kg. Right knee swelling is calculated as a ratio of left kneeswelling, and the R/L knee swelling ratio plotted against time to giveArea Under the Curve (AUC) graphs for control and treatment groups. Thepercentage inhibition of the individual treatment group AUCs arecalculated vs the control group AUC (0% inhibition) using an Excelspreadsheet.

[0089] On day 14, the mice are sacrificed by CO₂ inhalation and theright and left knees removed and processed for undecalcified histologyusing a Histodur plastic embedding method (Leica AG, Germany). Sections(5 μm) from both the control and arthritic knees are cut on a RM 2165rotation microtome (Leica AG, Germany). After staining, the slides arenumber coded as left knee/right knee pairs from each animal and scoredin a blinded fashion for inflammatory cell infiltrate/hyperplasia, jointdamage/erosions and cartilage proteoglycan loss.

[0090] Illustrative of the invention, the compound of example 1 inhibitsknee swelling in the mouse antigen-induced arthritis model whenadministered at a dose range of about 0.3 to 30 mg/kg p.o., and alsosignificantly reduces the histological damage to the knee.

[0091] The LFA-1 antagonists of the invention are, useful in thetreatment and/or prevention of diseases or disorders mediated byLFA-1/ICAM-1, ICAM-2 or ICAM-3 interactions, e.g., ischemia/reperfusioninjury, e.g., myocardial infarction, stroke, gut ischemia, renal failureor hemorrhagic shock, acute or chronic rejection of organ or tissueallo- or xenografts, infection diseases such as septic shock, adultrespiratory distress syndrome, or traumatic shock. They are also usefulin the treatment and/or prevention of acute or chronic inflammatorydiseases or disorders or autoimmune diseases, e.g., rheumatoidarthritis, systemic lupus erythematosus, hashimoto's thyroidis, multiplesclerosis, myasthenia gravis, diabetes type I and uveitis, cutaneousmanifestations of immunologically-mediated illnesses, inflammatory andhyperproliferative skin diseases (such as psoriasis, atopic dermatitis,alopecia aerata, allergic contact dermatitis, irritant contactdermatitis and further eczematous dermatitises, seborrhoeic dermatitis,lichen planus, pemphigus, bullous pemphigoid, epidermolysis bullosa,urticaria, angioedemas, vasculitides, erythema multiforme, cutaneouseosinophilias, lupus erythematosus, acne, granuloma annulare, pyodermagangrenosum, sun burns or toxic epidermal necrolysis), inflammatorybowel disease, ophthalmic inflammatory diseases or immune-mediatedconditions of the eye, such as auto-immune diseases, e.g., chronickeratitis, allergic conditions, e.g., vernal conjunctivitis,inflammatory conditions due to ocular surgery, e.g., keratoplasty. Forthe above uses the required dosage will of course vary depending on themode of administration, the particular condition to be treated and theeffect desired, in general, systemically at daily dosages of from about0.1 to about 30 mg/kg body weight. An indicated daily dosage in thelarger mammal is in the range from about 1 mg to about 200 mg,conveniently administered, e.g., in divided doses up to four times a dayor in sustained release form.

[0092] For topical use, administration of a 1%-3% concentration ofactive substance several times daily, e.g., 2 to 5 times daily, can beused.

[0093] The compounds of the invention may be administered systemicallyor topically, by any conventional route, in particular enterally, e.g.,orally, e.g., in the form of tablets or capsules, topically, e.g., inthe form of lotions, gels, ointments or creams, or in a nasal or asuppository form. Percutaneous administration via patches or otherdelivery systems may also be a possible route for prevention ortreatment of above diseases.

[0094] Pharmaceutical compositions comprising a compound of theinvention in association with at least one pharmaceutical acceptablecarrier or diluent may be manufactured in conventional manner by mixingwith a pharmaceutically acceptable carrier or diluent. Unit dosage formscontain, for example, from about 1 mg to about 100 mg of activesubstance.

[0095] Topical administration is, e.g., to the skin. A further form oftopical administration is to the eye.

[0096] The compounds of the invention may be administered in free formor in pharmaceutically acceptable salt form, e.g., as indicated above.Such salts may be prepared in conventional manner and exhibit the sameorder of activity as the free compounds.

[0097] In accordance with the foregoing the present invention furtherprovides:

[0098] (a) a method of antagonizing LFA-1 and inhibiting LFA-1/ICAM-1,ICAM-2 or ICAM-3 binding in a mammal in need thereof which comprisesadministering to a said subject an effective amount of a compound offormula I, II, Ia or IIa as defined herein, or a pharmaceuticallyacceptable salt thereof;

[0099] (b) a method for preventing or treating disorders or diseasesmediated by LFA-1/ICAM-1, ICAM-2 or ICAM-3 interactions, such asindicated above, e.g., rheumatoid arthritis, multiple sclerosis in amammal in need of such treatment, which method comprises administeringto said subject an effective amount of a compound of formula I, II Ia orIIa as defined herein, or a pharmaceutically acceptable salt thereof;

[0100] (c) a method for preventing or treating inflammatory diseases ordisorders (acute or chronic) or autoimmune diseases, e.g., as indicatedabove, in a mammal in need of such treatment, which method comprisesadministering to said subject an effective amount of a compound offormula I, II, Ia or IIa as defined herein, or a pharmaceuticallyacceptable salt thereof;

[0101] (d) a method of treating arthritis, in particular rheumatoidarthritis, in a mammal in need of such treatment, which method comprisesadministering to said subject an effective amount of a compound offormula I, II, Ia or IIa as defined herein, or a pharmaceuticallyacceptable salt thereof;

[0102] (e) a method of treating inflammatory and hyperproliferative skindisorders such as psoriasis, eczema and dermatitis, in particularallergic dermatitis such as allergic contact dermatitis and atopicdermatitis, which method comprises administering to a mammal in needthereof an effective amount of a compound of formula I, II, Ia or IIa asdefind herein, or a pharmaceutically acceptable salt thereof;

[0103] (f) a pharmaceutical composition for use in antagonizing LFA-1activity comprising a compound of formula I, II, Ia or IIa as definedherein in free form or pharmaceutically acceptable salt form inassociation with a pharmaceutically acceptable diluent or carriertherefor.

[0104] The compounds of the invention may be administered as the soleactive ingredient or together with other drugs in immunomodulatingregimens or other anti-inflammatory agents for the treatment orprevention of allo- or xenograft acute or chronic rejection orinflammatory or autoimmune disorders. For example, they may be used incombination with cyclosporins, rapamycins or ascomycins, or theirimmunosuppressive analogs, e.g., cyclosporin A, cyclosporin G, FK-506,ABT-281, ASM 981, rapamycin, 40-O-(2-hydroxy)ethyl-rapamycin, etc.;corticosteroids; cyclophosphamide; azathioprene; methotrexate; FTY 720;leflunomide; mizoribine; mycophenolic acid; mycophenolate mofetil;15-deoxyspergualine; immunosuppressive monoclonal antibodies, e.g.,monoclonal antibodies to leukocyte receptors, e.g., MHC, CD2, CD3, CD4,CD7, CD25, CD28, B7, CD40, CD45, or CD58 or their ligands otherimmunomodulatory compounds, e.g., CTLA4Ig, or other adhesion moleculeinhibitors, e.g., mAbs or low molecular weight inhibitors includingSelectin antagonists and VLA-4 antagonists. For the treatment ofarthritis, they can be used in combination, with, e.g., non-steroidalantiarthritic drugs, such as diclofenac, naproxen and ibuprofen, COX-2inhibitors such as rofecoxib, celecoxib, etoricoxib, valdecoxib,parecoxib, tiracoxib, COX-189, and steroids such as dexamethasone, andother agents, such as methotrexate, gold salts, d-penicillamine andcyclosporin A.

[0105] Where the compounds of formula I are administered in conjunctionwith other immunosuppressive/immunomodulatory or anti-inflammatorytherapy, e.g., for preventing or treating chronic rejection or arthritisas hereinabove specified, dosages of the co-administeredimmunosuppressant, immunomodulatory or anti-inflammatory compound willof course vary depending on the type of co-drug employed, e.g., whetherit is a steroid or a cyclosporin, on the specific drug employed, on thecondition being treated and so forth. In accordance with the foregoingthe present invention provides in a yet further aspect:

[0106] (a) a method of use as defined above comprisingco-administration, e.g., concomitantly or in sequence, of atherapeutically effective amount of a compound of formula I, Ia, II orIIa in free form or in pharmaceutically acceptable salt form, and asecond drug substance, said second drug substance being animmunosuppressant, immunomodulatory or anti-inflammatory drug, e.g., asindicated above; and

[0107] (b) a therapeutic combination, e.g., a kit, for use in any methodof use as defined above, comprising a pharmaceutical compositioncontaining a compound of formula I, II, Ia or IIa in free form or inpharmaceutically acceptable salt form, with at least one pharmaceuticalcomposition comprising an immunosuppressant, immunomodulatory oranti-inflammatory drug. The kit may include instructions for itsadministration.

EXAMPLES

[0108] The following examples show representative compounds encompassedby this invention and their synthesis. However, it should be clearlyunderstood that it is for purposes of illustration only.

Example 1 1H-1,4-Diazepine-1-acetamide,hexahydro-N-methyl-3-(2-methylpropyl)-α-[(2-naphthalenyl)methyl]-2,5-dioxo-4-[(3-quinolinyl)methyl]-,(αS, 3S)-

[0109]

[0110] N-t Boc-L-3-(2-naphthyl)alanine (5.0 g, 15.85 mmol) is added toDMF (50 ml) at room temperature. 1-Hydroxybenzotriazole (HOBT) (4.29 g,31.70 mmol) and 1,3-dicyclohexylcarbodiimide (DCCI) (9.81 g, 47.55 mmol)are added and the mixture is stirred for 5 min. 4-Methylmorpholine (NMM)(1.74 ml) and methylamine (16.0 ml) in THF are added and the mixture isstirred for 16 h. The white precipitate is collected and washed withDMF, (2×35 ml) and discarded. The remaining reaction mixture plusfiltrates is added to 800 ml 5% NaHCO₃. The white resulting precipitateis collected and washed with 2×25 mls H₂O. The precipitate is dissolvedin 100 ml CH₂Cl₂ and the mixture filtered to remove insoluble salts. Thefiltrate is washed with brine and dried over anhydrous magnesiumsulfate. The solvent is then removed to give product as a white solid,mp: 161-165° C.

[0111] Step 2

[0112] Product from step 1 (3.1 g, 9.44 mmol) is added to CH₂Cl₂ (60ml). At room temperature (r.t.), anhydrous HCl gas is bubbled throughthe reaction mixture for 1 hour (h). The volume of the reaction mixtureis reduced by half and the remainder is added to 100 ml diethyl etherwith rapid agitation. A white solid forms, is filtered off, washed with2×50 ml ether and dried to give the desired product, mp: 179-181° C.

[0113] Step 3

[0114] Product from step 2 (2.5 g, 9.44 mmol) is added to MeOH (50 ml)at room temperature. N,N-Diisopropylethylamine (DIEA) (3.28 ml, 18.9mmol) is added and the mixture is stirred for 5 min. Methyl acrylate(1.5 ml) is then added and the mixture is stirred overnight. Additionalmethyl acrylate (0.6 ml) is added and the mixture stirred for a total of48 h. The solvent is evaporated and the crude product is chromatographedusing 1% MeOH in CH₂Cl₂ to give a white solid, mp: 92-94° C.

[0115] Step 4

[0116] Product from step 3 (2.95 g, 9.38 mmol) is added to DMF (100 ml)at room temperature. N-t-Boc-L-leucine (2.2 g, 9.38 mmol) and DIEA (1.6ml, 9.38 mmol) are added and the mixture is stirred for 5 min. Thereaction is cooled to 0° C.,[O-(7-azabenzotriazol-1-yl)-1,1,3,3-tetramethyluroniumhexafluorophosphate] (HATU) (2.0 g, 9.38 mmol) is added and the reactionis allowed to warm to room temperature. After 24 h, the reaction iscooled to 0° C., N-t-Boc-L-leucine (4.3 g, 18.76 mmol), DIEA (3.3 ml)and HATU (7.6 g) are added. After a total of 48 h, solvent is removedand residue is dissolved in ethyl acetate. The solution is washed with2×100 ml H₂O, 3×50 ml 5% NaHCO₃, 3×200 ml 10% Na₂CO₃ and dried. Thesolvent is removed and the residue chromatographed using 1.5% to 2%CH₃OH in CH₂Cl₂, obtaining product as a pale yellow foam, mp: 52-56° C.

[0117] Step 5

[0118] Product from step 4 (3.5 g, 6.63 mmol) is added to CH₂Cl₂ (100ml) at room temperature. HCl gas is bubbled through the reaction mixturefor 1 h. Part of the solvent (70 ml) is removed, and diethyl ether isadded to form a white precipitate. The precipitate is filtered off,washed with ether and dried to give desired product, mp: 175-180° C.dec.

[0119] Step 6

[0120] Product from step 5 (1.82 g, 3.92 mmol) is added to THF (40 ml)at room temperature. DIEA (0.7 ml, 3.92 mmol) is added, and the mixtureis stirred for 10 min. Quinoline-3-carboxaldehyde (0.597 g, 3.80 mmol)is added, and the mixture stirred for 10 min. Sodiumtriacetoxyborohydride (1.25 g, 5.88 mmol) is added followed by aceticacid (2.0 ml). After 2 h, 100 ml saturated NaHCO₃ is added to thereaction mixture which is then extracted with diethyl ether. The etherextract is washed with brine, dried, and evaporated to dryness. Theresidue is chromatographed using 1% to 5% MeOH in CH₂Cl₂ to give thedesired product as a white foam, mp: 48-54° C.

[0121] Step 7

[0122] Product from step 6 (1.99 g, 3.50 mmol) is added to THF (15 ml)at room temperature. A solution of lithium hydroxide monohydrate (0.441g, 10.5 mmol) in H₂O(5 ml) is added dropwise over 5 min. After 2 h,solvent is removed, residue is washed with 3×50 ml hexane and dissolvedin H₂O. The solution is acidified with 1N HCl, neutralized withsaturated NaHCO₃ and extracted with CH₂Cl₂. The extracts are dried andevaporated to dryness to give the desired product as a white foam, mp:86-97° C.

[0123] Step 8

[0124] Product from step 7 (1.88 g, 3.39 mmol) is added to DMF (40 ml)at room temperature. DIEA (0.61 ml, 3.50 mmol) is added and the mixtureis stirred for 5 min. The reaction mixture is cooled to 0° C. and HATU(1.33 g, 3.50 mmol) is added. After 1 h at 0° C., the solvent isremoved, the residue is dissolved in CH₂Cl₂ and the solution washed with1×200 ml H₂O, 1×150 ml brine, and dried. The solvent is removed and theresidue is chromatographed using 1.5% to 3.5% MeOH in CH₂Cl₂, to givethe desired title product, mp: 125-127° C.

Example 2 1H-1,4-Diazepine-1-acetamide,hexahydro-N-methyl-3-(2-methylpropyl)-α-[(2-naphthalenyl)methyl]-2,5-dioxo-4-[(6-quinolinyl)methyl]-,(αS, 3S)-

[0125]

[0126] Product from step 5 in Example 1 (100 mg, 0.234 mmol) isdissolved in DMF (5 ml) at room temperature. DIEA (0.041 ml, 0.234 mmol)is added and the mixture is stirred for 5 min. 6-Bromomethylquinoline(52 mg, 0.234 mmol) is then added and the mixture is stirred for 48 h.Concentration and purification of the residue by chromatography using 2%to 3% MeOH in CH₂Cl₂ gives the desired intermediate as a white solid(MS: MH+569).

[0127] Further steps are carried out essentially as described in Example1 to yield the title compound as a solid, mp: 102-105° C.

Example 3 1H-1,4-Diazepine-1-acetamide,4-[(4-bromophenyl)methyl]hexahydro-N-methyl-3-(2-methylpropyl)-α-[(2-naphthalenyl)methyl]-2,5-dioxo-,αS, 3S)-

[0128]

[0129] The title compound is prepared following essentially theprocedures of Example 2. The product is isolated as a solid, mp:105-108° C.

Example 4 1H-1,4-Diazepine-1-acetamide,hexahydro-N-methyl-3-(2-methylpropyl)-α-[(2-naphthalenyl)methyl]-2,5-dioxo-4-[3-(4-pyridinyl)-2-propenyl]-,αS, 3S)-

[0130]

[0131] The title compound is prepared using essentially the proceduresof Example 1. The product is isolated as a solid, mp: 68-72° C.

Example 5 1H-1,4-Diazepine-1-acetamide,4-[(4-dimethylaminophenyl)methyl]hexahydro-N-methyl-3-(2-methylpropyl)-α-[(2-naphthalenyl)methyl]-2,5-dioxo-,αS, 3S)-

[0132]

[0133] The title compound is prepared following essentially theprocedures of Example 1. The product is isolated as a solid, mp: 92-94°C.

Example 6 1H-1,4-Diazepine-1,4-diacetamide,hexahydro-N¹-methyl-3-(2-methylpropyl)-α-[(2-naphthalenyl)methyl]-2,5-dioxo-N⁴-(4-pyridinyl)]-,αS, 3S)-

[0134]

[0135] The title compound is prepared following essentially theprocedures of Example 2. The product is isolated as foam, mp: MS(ESI):MH+530.

Example 7 1H-1,4-Diazepine-1-acetamide,hexahydro-4-[(4-hydroxy-3-methoxyphenyl)methyl]-N-methyl-3-(2-methylpropyl)-α-[(2-naphthalenyl)methyl]-2,5-dioxo-,αS, 3S)-

[0136]

[0137] The title compound is prepared following essentially theprocedures of Example 1. The product is isolated as a solid, mp: 67-69°C.

Examples 8-13

[0138] The following compounds are prepared according to the methodspreviously described.

Exam- m.p. (° C.) ple R₂ R₃ or MH⁺ 8 4-hydroxy-3-methoxybenzyl—CH₂CONHCH₃ MH⁺ = 547 9 3-quinolinylethyl 2-methylpropyl 60-67 10 benzyl2-methylpropyl 57-62 11 3-benzyloxybenzyl 2-methylpropyl 57-63 123-hydroxybenzyl 2-methylpropyl 78-82 13 4-pyridylmethyl 2-methylpropyl63-70

Example 14 1H-1,4-Diazepine-1-acetamide,hexahydro-N-methyl-3-(2-methylpropyl)-α-[(2-naphthalenyl)methyl]-2,5-dioxo-4-[(3-quinolinyl)methyl]-,αR, 3R)-

[0139]

[0140] The title compound is prepared essentially as described inExample 2. The product is isolated as a solid, mp: 43-54° C.

Example 15 1H-1,4-Diazepine-1-acetamide,hexahydro-N-methyl-3-(2-methylpropyl)-α-[(2-naphthalenyl)methyl]-2,5-dioxo-4-[(3-quinolinyl)methyl]-, αS, 3R)-

[0141]

[0142] The title compound is prepared essentially as described inExample 2. The product is isolated as a solid, mp: 53-59° C.

Example 16 1H-1,4-Diazepine-1-acetamide,hexahydro-N-methyl-3-(2-methylpropyl)-α(-[(2-naphthalenyl)methyl]-2,5-dioxo-4-[(3-quinolinyl)methyl]-, (αR, 3S)-

[0143]

[0144] The title compound is prepared essentially as described inExample 2. The product is isolated as a solid; mp: 38-44° C.

Example 17 1-Piperazineacetamide,hexahydro-N-methyl-3-(2-methylpropyl)-α-[(2-naphthalenyl)methyl]-2,5-dioxo-4-[(6-quinolinyl)methyl]-,αS, 3S)-

[0145]

[0146] L-leucine t-butyl ester hydrochloride (287 mg, 1.3 mmol) isdissolved in DMF (4 ml), and DIEA (0.475 ml, 2.1 mmol) is addeddropwise. A solution of 4-bromomethylquinoline (200 mg, 0.9 mmol) in DMF(4 ml) is added dropwise, the mixture is stirred for 16 h at rt,concentrated to dryness and chromatographed using 2% CH₃OH/CH₂Cl₂ togive a yellow oil.

[0147] To a solution of product from step 1 (40 mg, 0.12 mmol) in DMF (2ml) at 0° C. is added dropwise a premixed solution of chloroacetic acid(19.6 mg, 0.21 mmol) and diisopropylcarbodiimide (DIPCI) (0.034 ml, 0.22mmol) in DMF (1 ml). The reaction mixture is stirred and allowed toreach room temperature. After 2 h, the reaction mixture is added tobrine, the mixture is extracted with ethyl acetate, the extract is driedand concentrated to dryness. The crude product is used directly in thenext step.

[0148] Crude product from step 2 (0.12 mmol) is dissolved in DMF (3 ml).L-3-(2-naphthyl)alanine N-methylamide (the free base of product fromstep 2 of example 1) (35 mg, 0.12 mmol) is added followed by DIEA (0.031ml, 0.18 mmol). The reaction mixture is stirred at 40° C. for 16 h andthen at 70° C. for 2 h, concentrated to dryness and chromatographedusing 7% CH₃OH/CH₂Cl₂ to obtain crude product of above structure.

[0149] To a solution of product from step 3 (25.8 mg, 0.043 mmol) inCH₂Cl₂ (3 ml) at rt is added TFA (0.8 ml). After 3 h, the reactionmixture is concentrated and the crude product is used directly in thenext step.

[0150] To a solution of the crude product from step 4 (0.043 mmol) inDMF (2 ml) is added DIEA (0.03 ml, 0.172 mmol) or enough to render thesolution basic. The solution is cooled to 0° C. and HATU (17 mg, 0.044mmol) is added. The reaction mixture is stirred at rt for 4 h,concentrated to dryness and purified by reverse phase chromatography(CH₃CN and 0.1% TFA/water) to give product as TFA salt, mp: 98-102° C.

Example 18 1-Piperazineacetamide,4-[(4-bromophenyl)methyl]hexahydro-N-methyl-3-(2-methylpropyl)-α-[(2-naphthalenyl)methyl]-2,5-dioxo-,αS, 3S)-

[0151]

[0152] The title compound is prepared following essentially theprocedures of Example 17. The product is isolated as a solid, mp:107-111° C.

Example 19 1-Piperazineacetamide,hexahydro-N-methyl-3-(2-methylpropyl)-α-[(2-naphthalenyl)methyl]-2,5-dioxo-4-[4-[(3-pyridinyl)phenyl]methyl]-,(αS, 3S)-

[0153]

[0154] The title compound is prepared from Example 18 by Stille couplingwith 3-(tri-n-butylstannyl)pyridine using a reported procedure (S.Wattanasin, Synth. Commn., 18, 1919, 1988). The product is isolated assolid, mp: 50-53° C.

What is claimed is:
 1. A compound of the formula I

wherein R is carboxy, esterified carboxy or amidated carboxy; R₁ and R₃are independently lower alkyl, (hydroxy, lower alkoxy, amino, acylamino,mono- or di-lower alkylamino or mercapto)-lower alkyl, loweralkyl-(thio, sulfinyl or sulfonyl)-lower alkyl, cycloalkyl, aryl,biaryl, (cycloalkyl, aryl or biaryl)-lower alkyl, or (carboxy,esterified carboxy or amidated carboxy)-lower alkyl; R₂ is hydrogen,lower alkyl, cycloalkyl, aryl, biaryl, arylaminocarbonyl, or aryl-(oxy,thio or amino); n is 1 or 2; Y is lower-alkylene or lower alkenylene; ora pharmaceutically acceptable salt thereof.
 2. A compound according toclaim 1 wherein n is
 2. 3. A compound according to claim 2 wherein R iscarboxy derivatized in form of a pharmaceutically acceptable amide; R₁is aryl-lower alkyl; R₂ is aryl; R₃ is lower alkyl, aryl, orcycloalkyl-lower alkyl; and Y is C₁-C₄-alkylene or C₂-C₄-alkenylene; ora pharmaceutically acceptable salt thereof.
 4. A compound of the formulaII

wherein R₁′ is bicyclic aryl-lower alkyl; R₂′ is bicyclic aryl; R₃′ ismonocyclic aryl or lower alkyl; R₄ is hydrogen or lower alkyl; n is 1 or2; Y is C₁₋₄-alkylene; or a pharmaceutically acceptable salt thereof. 5.A compound according to claim 4 of formula II wherein R₁ ′ isnaphthyl-lower alkyl or quinolinyl-lower alkyl; R₂′ is quinolinyl; Y ismethylene; and n is 2; or pharmaceutically acceptable salt thereof.
 6. Acompound according to claim 1 the formula Ia

wherein n, Y, R, R₁, R₂ and R₃ have meaning as defined in said claim; ora pharmaceutically acceptable salt thereof.
 7. A compound according toclaim 4 of the formula IIa

wherein n, Y, R₁′, R₂′, R₃′ and R₄ have meaning as defined in saidclaim; or a pharmaceutically acceptable salt thereof.
 8. A compoundaccording to claim 7 of formula IIa, wherein R₁ is 2-naphthylmethyl; R₂′is 3-quinolinyl; R₃′ is isobutyl; R₄ is methyl; Y is CH₂; and n is 2; ora pharmaceutically acceptable salt thereof.
 9. A process for thepreparation of a compound according to claim 1 of formula I whichprocess comprises: (a) cyclizing a compound of formula III

or a reactive functional derivative thereof, wherein R, R₁-R_(3,) Y andn have meaning as defined in claim 1; or (b) cyclizing a compound offormula IV

or a reactive functional derivative thereof, wherein R, R₁-R₃, Y and nhave meaning as defined in claim 1; and if desired converting theresulting compound of formula I into a pharmaceutically acceptable saltthereof.
 10. A pharmaceutical composition comprising a compoundaccording to claim 1 in combination with a pharmaceutically acceptablecarrier.
 11. A method of antagonizing LFA-1 activity in mammals whichcomprises administering to a mammal in need thereof an effective amountof a compound according to claim
 1. 12. A method of treatinginflammatory or autoimmune disorders in mammals which comprisesadministering to a mammal in need thereof an effective amount of acompound according to claim
 1. 13. A method according to claim 12 oftreating arthritis.
 14. A method according to claim 12 of treatingrheumatoid arthritis.
 15. A method of treating hyperproliferative andinflammatory skin disorders in mammals which comprises administering toa mammal in need thereof an effective amount of a compound according toclaim
 1. 16. A method according to claim 15 of treating psoriasis,eczema and dermatitis.
 17. A method according to claim 15 of treatingallergic contact dermatitis and atopic dermatitis.